Division of Pharmacognosy, Section Metabolomics, Institute of Biology, Leiden University, Einsteinweg, Leiden, The Netherlands.
Nat Protoc. 2010 Mar;5(3):536-49. doi: 10.1038/nprot.2009.237. Epub 2010 Feb 25.
Nuclear magnetic resonance (NMR)-based metabolomics has many applications in plant science. Metabolomics can be used in functional genomics and to differentiate plants from different origin, or after different treatments. In this protocol, the following steps of plant metabolomics using NMR spectroscopy are described: sample preparation (freeze drying followed by extraction by ultrasonication with 1:1 CD(3)OD:KH(2)PO(4) buffer in D(2)O), NMR analysis (standard (1)H, J-resolved, (1)H-(1)H correlation spectroscopy (COSY) and heteronuclear multiple bond correlation (HMBC)) and chemometric methods. The main advantage of NMR metabolomic analysis is the possibility of identifying metabolites by comparing NMR data with references or by structure elucidation using two-dimensional NMR. This protocol is particularly suited for the analysis of secondary metabolites such as phenolic compounds (usually abundant in plants), and for primary metabolites (e.g., sugars and amino acids). This procedure is rapid; it takes not more than 30 min for sample preparation (multiple parallel) and a further 10 min for NMR spectrum acquisition.
基于核磁共振（NMR）的代谢组学在植物科学中有许多应用。代谢组学可用于功能基因组学，并可用于区分不同来源或经过不同处理的植物。在本方案中，描述了使用 NMR 光谱进行植物代谢组学的以下步骤：样品制备（冷冻干燥，然后用 1:1 CD(3)OD:KH(2)PO(4)缓冲液在 D(2)O 中进行超声提取），NMR 分析（标准 (1)H、J 分辨、(1)H-(1)H 相关光谱 (COSY) 和异核多键相关 (HMBC))和化学计量方法。NMR 代谢组学分析的主要优势在于可以通过将 NMR 数据与参考数据进行比较，或者通过二维 NMR 进行结构解析来识别代谢物。该方案特别适合分析次级代谢产物，如酚类化合物（通常在植物中含量丰富），以及初级代谢产物（如糖和氨基酸）。该程序快速；样品制备（多平行）不超过 30 分钟，NMR 光谱采集进一步需要 10 分钟。
